LOCUS       sequence-289984-        8689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot1iso1_4-1317.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8689)
  AUTHORS   Bawankar P, Loh B, Wohlbold L, Schmidt S, Izaurralde E
  TITLE     NOT10 and C2orf29/NOT11 form a conserved module of the CCR4-NOT 
            complex that docks onto the NOT1 N-terminal domain.
  JOURNAL   RNA Biol. 2013 Feb;10(2):228-44. doi: 10.4161/rna.23018. Epub 2013 
            Jan 9.
  PUBMED    23303381
REFERENCE   2  (bases 1 to 8689)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.4161/rna.23018";
            journalName: "RNA Biol"; date: "2013-02"; volume: "10"; issue: "2"; 
            pages: "228-44"
COMMENT     Sequence Label: pT7-EGFP-C1-HsNot1iso1_4-1317_S
FEATURES             Location/Qualifiers
     source          1..8689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(710..731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(971..990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1318..1339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5477..5598
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5514..5533)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5568..5587
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5605..6060)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5742..5763)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5954..5973
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6087..6191
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6193..6550
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6193..6384
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6216..6236)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6401..6536
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6585..7379
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6585..7379
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6639..6658)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7249..7268
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7567..7586)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7611..7658
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7987..8575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8476..8495
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      661 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1381 gtccggactc agatctcgag ctgactcgct ctcgctggcc ttgtctcaaa tcagctacct
     1441 ggtggacaat ttaaccaaga aaaattaccg agccagccag caggaaatac agcatattgt
     1501 gaatcggcac ggtcctgagg cagacaggca tttattacgc tgcctatttt cgcatgtgga
     1561 tttcagtggc gatggtaaaa gcagtggcaa agatttccat cagactcagt ttctgattca
     1621 ggagtgtgcg ttgctgatta caaagccaaa ttttatctcg acgctgtcct atgccattga
     1681 taatccattg cactatcaga agagtttaaa gcctgcaccc cacttatttg cccagctgag
     1741 taaagtgctc aaattaagca aagtacaaga ggtaattttt ggccttgccc tgttgaattc
     1801 ttccagctca gatcttagag gtttcgctgc ccagtttatc aaacagaagc ttccagatct
     1861 tctgcgttct tacattgacg cagacgtcag tggaaatcaa gaaggtggct tccaagatat
     1921 agcaatagag gtcctacacc tcctcctctc ccatctcctc tttgggcaga agggagcctt
     1981 tggagttgga caagaacaga tagacgcttt tcttaagacg ctgcgcagag attttcccca
     2041 agaacgctgt cccgtggtgc tcgcaccact tttataccct gaaaaacggg acattctaat
     2101 ggacaggatc ctgcctgatt ccggaggggt agctaaaacc atgatggaga gctctttggc
     2161 tgatttcatg caagaagtag gctatggctt ttgtgcaagt attgaagaat gtcgcaatat
     2221 aatcgtgcag tttggtgttc gggaggtcac agctgcccag gttgcaaggg ttttgggaat
     2281 gatggctcga actcattcag gattaacaga tggcattcca ttacagagta tttctgctcc
     2341 gggcagtggg atctggagtg atgggaaaga taaaagtgat ggagcacagg cacacacatg
     2401 gaatgtagaa gtcttgattg acgttcttaa agaactgaat ccaagtttga atttcaagga
     2461 agtaacttat gaactggacc atcctggatt tcaaattcgt gacagtaaag gacttcataa
     2521 tgtggtttat ggcattcaga ggggtttggg tatggaagtg ttcccagtag acctcatata
     2581 tagaccttgg aaacatgctg aaggccagct ctccttcatt caacattccc ttataaatcc
     2641 agagatcttc tgttttgctg actatccctg tcatactgtt gccactgata ttctgaaagc
     2701 accaccagag gatgacaatc gagaaattgc cacatggaag agcttggatt tgattgaatc
     2761 tctgctgagg cttgcagagg ttgggcagta tgagcaagtc aaacagctct tcagcttccc
     2821 tatcaaacac tgtccagaca tgctggtatt ggccttacta caaattaaca cctcttggca
     2881 taccttgcgc catgaactta tctccactct gatgccaatt ttccttggaa accatcctaa
     2941 ctcagctatt attttgcact atgcatggca tgggcaggga cagtctccct caattcgcca
     3001 acttatcatg catgcaatgg cagaatggta catgagaggg gagcagtatg atcaggccaa
     3061 attgtctcga atacttgatg tggcccagga cttgaaggcc ttgtcaatgc tgctaaatgg
     3121 tactccattt gcctttgtta ttgaccttgc tgcacttgct tcacgtcgtg aatacctcaa
     3181 acttgataag tggctcacag ataaaattcg agagcatggg gagcctttta tccaggcgtg
     3241 tatgactttt ttaaagagac ggtgtccttc tattttgggc ggacttgccc cagaaaaaga
     3301 ccagcccaaa agtgctcaac ttcctccaga aactttggcg acaatgttgg cctgtctgca
     3361 agcttgtgca gggagtgttt ctcaggagct atcagaaact atcctcacca tggtagccaa
     3421 ttgcagtaat gttatgaata aggccagaca accaccacct ggagttatgc caaaaggacg
     3481 tcctcctagt gctagcagct tagatgccat ttctcctgtt cagattgacc ctcttgctgg
     3541 aatgacatct cttagtatag gtggttcagc tgcccctcac acccagagta tgcagggttt
     3601 tcctccaaat ttgggttctg cattcagtac ccctcagtca ccagcaaaag catttccacc
     3661 cctttcaacc cccaatcaga ccactgcatt cagtggtatt ggaggacttt catcacagct
     3721 tccagtaggt ggtcttggca caggcagcct gactggtata ggaactggtg ctcttggact
     3781 ccctgcagtg aataacgacc cttttgtaca gaggaaactg ggcacctctg gactgaatca
     3841 gcctacattc cagcagacgg acttgtctca ggtgtggcca gaggcaaacc agcactttag
     3901 taaagagata gatgatgaag caaacagcta tttccagcga atatataatc atccaccaca
     3961 tccaaccatg tctgttgatg aggtattaga aatgctgcag agatttaaag actctactat
     4021 aaagagggaa cgagaagtat ttaactgtat gctaaggaac ttgtttgaag aatatcgttt
     4081 ttttccccag tatcctgata aagagttaca tataacagcc tgcctatttg gtggtataat
     4141 tgagaaagga ctggtcactt acatggcact aggtctggct ctacgatatg ttcttgaagc
     4201 cttacgcaag ccttttggat ccaaaatgta ttatttcggg attgctgcac tagatagatt
     4261 taaaaacaga ttgaaggact atccccagta ttgtcagcat ttggcttcta tcagtcactt
     4321 tatgcaattt ccacatcatt tacaggagta tattgagtat ggacagcagt ctagagatcc
     4381 tcctgtgaaa atgcaaggct ctatcacaac ccctggaagt attgcactgg ctcaggccca
     4441 ggctcaggcc caggttccag caaaagctcc tcttgctggt caagttagca ctatggtaac
     4501 cacctcaaca actaccactg ttgctaaaac ggttacggtc accaggccaa ctggagtcag
     4561 ctttaagaaa gatgtgccac cttctattaa tactacaaat atagatacgt tgcttgtggc
     4621 cacagatcaa actgagagaa ttgtggagcc cccagaaaat atccaggaga aaattgcttt
     4681 tattttcaat aatctctcac agtcaaatat gacacaaaag gttgaagagc taaaggaaac
     4741 ggtgaaagaa gaatttatgc cttgggtttc acagtatctg gttatgaaga gagtcagtat
     4801 tgagccaaac tttcatagcc tgtattcaaa cttccttgac acgctgaaga atcctgaatt
     4861 taacaagatg gttctgaatg agacctacag aaacattaaa gtgctcctga cctctgataa
     4921 agctgcagcc aatttctcag atcgttcttt gctgaagaac ttgggacatt ggctaggaat
     4981 gatcacatta gctaaaaaca aacccatctt acacactgac ttggatgtga aatcattgct
     5041 gctagaggct tatgttaaag gacaacaaga attgctctat gtagtgccct ttgttgccaa
     5101 agtcttagaa tctagcattc gtagtgtggt ttttaggcca ccaaaccctt ggacaatggc
     5161 aattatgaat gtattagctg agctacatca ggagcatgac ttaaagttaa acttgaagtt
     5221 tgaaatcgag gttctctgca agaaccttgc attagacatc aatgagctaa aacctggaaa
     5281 cctcctaaag gataaagatc gcctgaagaa tttagatgag caactctctt aaggtaccgc
     5341 gggcccggga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     5401 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     5461 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5521 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5581 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     5641 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     5701 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     5761 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     5821 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     5881 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     5941 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     6001 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6061 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6121 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6181 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     6241 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6301 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6361 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     6421 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6481 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     6541 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     6601 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     6661 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     6721 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     6781 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     6841 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     6901 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     6961 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     7021 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     7081 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     7141 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     7201 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     7261 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     7321 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     7381 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     7441 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     7501 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     7561 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     7621 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     7681 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     7741 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     7801 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     7861 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7921 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7981 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     8041 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     8101 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     8161 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     8221 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     8281 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     8341 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     8401 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     8461 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     8521 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     8581 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     8641 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
